Mechanically patterned neuromuscular junctions-in-a-dish have improved functional maturation.

Abstract:

:Motor neuron (MN) diseases are progressive disorders resulting from degeneration of neuromuscular junctions (NMJs), which form the connection between MNs and muscle fibers. NMJ-in-a-dish models have been developed to examine human MN-associated dysfunction with disease; however such coculture models have randomly oriented myotubes with immature synapses that contract asynchronously. Mechanically patterned (MP) extracellular matrix with alternating soft and stiff stripes improves current NMJ-in-a-dish models by inducing both mouse and human myoblast durotaxis to stripes where they aligned, differentiated, and fused into patterned myotubes. Compared to conventional culture on rigid substrates or unpatterned hydrogels, MP substrates supported increased differentiation and fusion, significantly larger acetylcholine (ACh) receptor clusters, and increased expression of MuSK and Lrp4, two cell surface receptors required for NMJ formation. Robust contractions were observed when mouse myotubes were stimulated by ACh, with twitch duration and frequency most closely resembling those for mature muscle on MP substrates. Fused myotubes, when cocultured with MNs, were able to form even larger NMJs. Thus MP matrices produce more functionally active NMJs-in-a-dish, which could be used to elucidate disease pathology and facilitate drug discovery.

journal_name

Mol Biol Cell

authors

Happe CL,Tenerelli KP,Gromova AK,Kolb F,Engler AJ

doi

10.1091/mbc.E17-01-0046

subject

Has Abstract

pub_date

2017-07-07 00:00:00

pages

1950-1958

issue

14

eissn

1059-1524

issn

1939-4586

pii

mbc.E17-01-0046

journal_volume

28

pub_type

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